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Method, reagent and kit for malaria testing

a malaria and kit technology, applied in the field of malaria testing, can solve the problems of inability to easily carry out any of these test methods in endemic areas, inability to have sufficient knowledge and experience, and inability to be said to be practical as test methods, etc., to achieve significant increases in the amount of urinary l-fabp, increase of the amount of l-fabp in blood, and increase of the amount of l-fabp in urine

Active Publication Date: 2010-03-18
THE UNIV OF TOKYO +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0022]The present inventors carried out diligent research in order to solve the foregoing problems. Consequently, it was found that there is a correlation between malaria infection and the amount of the urinary L-FABP of a subject animal. Conventionally, it has been known that the amount of the L-FABP in blood is significantly increased in the case of liver disease, while the amount of the urinary L-FABP is not increased (Non-patent Document 4). In other words, an increase of the amount of the urinary L-FABP resulting from malaria infection is not relevant to the disclosure in Non-patent Document 3. Additionall

Problems solved by technology

However, in the method described above including microscopic observation of a smear preparation, each of the steps of: production of a smear preparation; fixation; staining; and drying must be carried out, and thus complicated operations, as well as sufficient knowledge and experience are required.
Moreover, the method disclosed in Patent Document 1 necessitates complicated operations, and special apparatus such as a flow cytometer is required.
Accordingly, it cannot be said that any of these test methods could be readily carried out in endemic areas of malaria.
However, in order to apply such findings to methods for malaria testing, L-FABP expression in hepatocytes have to be determined, and thus, they are not practical as test methods.
Therefore, it is also not practical as a test method in this respect.
Furthermore, the extent of infection with malaria cannot be known from this test method, in contrast to the methods including microscopic observation of a smear preparation, and the like.
Accordingly, no convenient method for malaria testing which focuses on a L-FABP has been known so far.

Method used

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  • Method, reagent and kit for malaria testing
  • Method, reagent and kit for malaria testing
  • Method, reagent and kit for malaria testing

Examples

Experimental program
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Effect test

example 1

Production of Transgenic Mouse into which Human L-FABP Gene is Introduced (hL-FABP-Tg mouse)

[0061]In order to produce transgenic mice into which a human L-FABP gene is introduced (hL-FABP-Tg mouse), male BCF1 mice no younger than 13 weeks old for infertile mating and natural mating, female ICR mice no younger than 10 weeks old for embryo transplantation and foster parent, male BDF1 mice no younger than 13 weeks old for mating, and female BDF1 mice no younger than 8 weeks old for ovum collection were used, respectively. Thus obtained transgenic mice (B6C3F1) were back crossed with BALB / cA mice to produce hL-FABP-Tg mice.

example 2

Amount of Urinary hL-FABP Excreted by hL-FABP-Tg Mouse Infected with P. berghei ANKA Strain

[0062]The P. berghei ANKA strain causes fatal infection in Balb / cA mice. In experimental infection, an intraerythrocytic protozoan parasite of P. berghei ANKA strain was used, and the hL-FABP-Tg mice (n=3) produced in Example 1 were used as subject animals. Additionally, in order to prevent influence by passive immunity, a Balb / cA RAG-2 knock out mouse was infected with the protozoan parasite of P. berghei ANKA strain, and used as a donor of parasitized erythrocytes for use in the experiment.

[0063]The hL-FABP-Tg mouse was intraperitoneally infected with 1×106 cells of parasitized fresh erythrocytes obtained from the infected RAG-2 knock out mouse. Following infection, the rate of parasitized erythrocytes in peripheral blood in the hL-FABP-Tg mouse was determined with Giemsa-stained blood smear preparations. The results are shown in FIG. 1.

[0064]In addition, spot urine was collected every day b...

example 3

Histopathological Analysis of Kidney in hL-FABP-Tg Mouse on Day 7 Following Infection with P. berghei ANKA Strain

[0067]The hL-FABP-Tg mouse on day 7 following infection with the P. berghei ANKA strain used in Example 2 was dissected for inspection, and histopathological analysis of the kidney was carried out by PAS staining. As a control, a hL-FABP-Tg mouse administered with cisplatin was used (Negishi K. et al., Kidney Int. 73 (12): 1374-1384(2008)). With respect to this control mouse, after a male 8- to 10-week old hL-FABP-Tg mouse was preliminarily fed for one week, 20 mg / kg of cisplatin dissolved in physiological saline (total solution: 60 mL / kg) was intraperitoneally administered once, and the mouse was dissected for inspection 72 hours later to carry out a histopathological analysis of the kidney by PAS staining. This control mouse exhibited an amount of hL-FABP excreted into urine of a similar level to the amount of excretion from the hL-FABP-Tg mouse on day 7 following infec...

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Abstract

The present invention provides a method for malaria testing which enables testing for the presence of malaria infection and the extent of the infection in a convenient manner; and a reagent or kit which can be used in the method. The method for malaria testing according to the present invention includes the step of detecting a liver-type fatty acid binding protein present in urine collected from a subject animal. The extent of infection is determined to be higher when a larger amount of the liver-type fatty acid binding protein is present in a test sample.

Description

[0001]This application is based on and claims the benefit of priority from Japanese Patent Application No. 2008-238680, filed on 17 Sep. 2008, the content of which is incorporated herein by reference.BACKGROUND OF THE INVENTION[0002]1. Field of the Invention[0003]The present invention relates to a method for malaria testing, and a reagent or kit for testing, which can be used in the test method.[0004]2. Related Art[0005]Malaria is an infectious disease caused by infection with a malaria parasite, which is mediated by mosquitoes belonging to the genus Anopheles. Malaria parasites (Plasmodium spp.) are protozoa belonging to Apicomplexa, the members of which are obligate intracellular parasites, in other words, they can proliferate only in cells. Protozoan parasites exhibit comparatively high host specificity, and there are four kinds of malaria parasites that utilize humans as a definitive host, i.e., falciparum malaria parasites (P. falciparum), tertian malaria parasites (P. vivax), ...

Claims

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Application Information

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IPC IPC(8): C12Q1/00
CPCG01N2333/445G01N33/56905
Inventor SUGAYA, TAKESHINOIRI, EISEIMATSUMOTO, YOSHITSUGU
Owner THE UNIV OF TOKYO